ABSTRACT
Objective To establish a method for the isolation and culture of bone marrow derived mesenchymal stem cells (MSCs) from patients with coronary artery disease and investigated its biological characteristics. Methods Bone marrow was extracted from patients' sternum when they were operated for coronary artery bypass graft. MSCs were separated from bone marrow by density gradient centrifugation in Percoll solution and seeded in culture medium supplemented with 10% fetal bovine serum. The morphology and growth characteristic of MSCs were observed in primary and passage generation culture with inverted microscope. The cell phenotype and cell cycle of MSCs were analyzed by flow cytometry. The growth curves were compared between different passages and ages. Results Most adherent MSCs showed spindle morphology whatever passages they were. The MSCs could be expanded for 15 passages. The proliferative potential of MSCs over the fifth passage or from old patients decreased gradually. The MSCs in primary and passage were positive for CD29 and CD44, but negative for CD34 and CD45. Analysis for cell cycle showed most MSCs in G_0/G_1 phase. Conclusion The method of isolation and culture of sternum bone marrow derived MSCs from patients with coronary artery disease is feasible, the MSCs have self-renewal capacity and might be ideal for cellular cardiomyoplasty.